Nuclear Receptor Coactivator 1 (NCOA1) (AA 761-863) antibody

Antigen: Nuclear Receptor Coactivator 1 (NCOA1)

Synonyms: SRC1, KAT13A, RIP160, F-SRC-1, bHLHe42, bHLHe74, MGC129719, MGC129720, NCOA1, si:dkey-5g7.6, si:dkey-34f16.6, SRA, SRAP, STRAA1, pp7684, MGC87674, Sra, Srap, Straa1, Strra1, AA959952, zgc:86613, SRA1, MGC128615, DKFZp459C083

Binding Site: AA 761-863

Clonality: Monoclonal (8)

Host: Mouse

Reactivity: Human

Conjugate: Un-conjugated

Application: Western Blotting (WB), BioImaging (BI)

Catalog no.: ABIN968842

Additional Information

Alternative name: SRC-1

Immunogen: Human SRC-1

Cross-Reactivity: Mouse (Murine), Rat (Rattus)

Format: Liquid

Isotype: IgG2a

Clone: 8

Description: Signal transduction via nuclear hormone receptors is important for cell growth and differentiation, development, and homeostasis. Nuclear hormone receptors are ligand-activated transcription factors that modulate target gene expression.These ligand/receptor complexes also interact with transcriptional coactivators which enhance ligand-dependent transcription. Various classes of coactivators have been identified, including SRC-1 and its related proteins, such as TIF-2/GRIP-1, RIP140, AIB-1, and TIF-1alpha and -1ß. SRC-1 is a member of the basic helix-loop-helix-PAS domain family. It contains an N-terminal basic-helix-loop-helix (bHLH), two N-terminal PAS domains, and a glutamine-rich region near the C-terminus. SRC-1 interacts with many different steroid receptors and recruitment of SRC-1 to promoter sites may facilitate opening of chromatin structure via its histone acetyltransferase activity, as well as stabilization of the preinitiation complex. ERK-2 can phosphorylate SRC-1 at threonine and serine sites in vitro. In addition, SRC-1 expression in cells stimulated with EGF enhances progesterone receptor-mediated activation of a target reporter gene. Thus, SRC-1 is an important transcriptional co-activator involved in steroid receptor signaling.

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.

Molecular Weight: 165 kDa

Comments:

Related Products: ABIN967389, ABIN968537

Synonyms: SRC1, KAT13A, RIP160, F-SRC-1, bHLHe42, bHLHe74, MGC129719, MGC129720, NCOA1, si:dkey-5g7.6, si:dkey-34f16.6, SRA, SRAP, STRAA1, pp7684, MGC87674, Sra, Srap, Straa1, Strra1, AA959952, zgc:86613, SRA1, MGC128615, DKFZp459C083

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Concentration: 250 µg/ml

Purity: Purified

Purification: Purified from tissue culture supernatant or ascites by affinity chromatography.

Buffer: Aqueous buffered solution containing BSA, glycerol.

Preservative: 0.09% Sodium azide.

Storage: Store undiluted at -20° C.

Restrictions: For Research Use only

Images

Western blot analysis of SRC-1 on a Jurkat lysate. Lane 1: 1:250, lane 2: 1:500, lane 3: 1:1000 dilution of the SRC-1 antibody.

Immunofluorescent staining of A549 (ATCC CCL-185) cells. Cells were seeded in a 96 well imaging plate at ~ 10 000 cells per well. After overnight incubation, cells were stained using the alcohol perm protocol and the anti-SRC-1 antibody. The second step reagent was FITC goat anti mouse Ig. Images were taken on a BD Pathway™ 855 Bioimager using a 20x objective. This antibody also stained HeLa (ATCC CCL-2)and U-2 OS (ATCC HTB-96) cells and worked with both the Triton™ X-100 and alcohol perm protocols.

Publications

Product: Oñate, Tsai, Tsai et al.: "Sequence and characterization of a coactivator for the steroid hormone receptor superfamily." in: Science (New York, N.Y.), Vol. 270, Issue 5240, pp. 1354-7, 1995 (PubMed).

Yao, Ku, Zhou et al.: "The nuclear hormone receptor coactivator SRC-1 is a specific target of p300." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 93, Issue 20, pp. 10626-31, 1996 (PubMed).

Spencer, Jenster, Burcin et al.: "Steroid receptor coactivator-1 is a histone acetyltransferase." in: Nature, Vol. 389, Issue 6647, pp. 194-8, 1997 (PubMed).

Rowan, Weigel, OMalley: "Phosphorylation of steroid receptor coactivator-1. Identification of the phosphorylation sites and phosphorylation through the mitogen-activated protein kinase pathway." in: The Journal of biological chemistry, Vol. 275, Issue 6, pp. 4475-83, 2000 (PubMed).